title: Immunosuppressive properties of mesenchymal stromal cell cultures derived from the limbus of human and rabbit corneas.
abstract: OBJECTIVE
  Mesenchymal stromal cells (MSCs) cultivated from the corneal limbus (L-MSCs) provide a potential source of cells for corneal repair. In the present study, we investigated the immunosuppressive properties of human L-MSCs and putative rabbit L-MSCs to develop an allogeneic therapy and animal model of L-MSC transplantation.
  
  
  METHODS
  MSC-like cultures were established from the limbal stroma of human and rabbit (New Zealand white) corneas using either serum-supplemented medium or a commercial serum-free MSC medium (MesenCult-XF Culture Kit; Stem Cell Technologies, Melbourne, Australia). L-MSC phenotype was examined by flow cytometry. The immunosuppressive properties of L-MSC cultures were assessed using mixed leukocyte reactions. L-MSC cultures were also tested for their ability to support colony formation by primary limbal epithelial (LE) cells.
  
  
  RESULTS
  Human L-MSC cultures were typically CD34⁻, CD45⁻ and HLA-DR⁻ and CD73⁺, CD90⁺, CD105⁺ and HLA-ABC⁺. High levels (>80%) of CD146 expression were observed for L-MSC cultures grown in serum-supplemented medium but not cultures grown in MesenCult-XF (approximately 1%). Rabbit L-MSCs were approximately 95% positive for major histocompatibility complex class I and expressed lower levels of major histocompatibility complex class II (approximately 10%), CD45 (approximately 20%), CD105 (approximately 60%) and CD90 (<10%). Human L-MSCs and rabbit L-MSCs suppressed human T-cell proliferation by up to 75%. Conversely, L-MSCs from either species stimulated a 2-fold to 3-fold increase in LE cell colony formation.
  
  
  CONCLUSIONS
  L-MSCs display immunosuppressive qualities in addition to their established non-immunogenic profile and stimulate LE cell growth in vitro across species boundaries. These results support the potential use of allogeneic L-MSCs in the treatment of corneal disorders and suggest that the rabbit would provide a useful pre-clinical model.
authors.affiliation: Queensland Eye Institute, South Brisbane, Queensland, Australia; School of Biomedical Sciences, Faculty of Health, Queensland University of Technology, Brisbane, Queensland, Australia; Mater Medical Research Institute, South Brisbane, Queensland, Australia. Electronic address: laura.bray@qut.edu.au.
authors.forename: Laura J
authors.initials: LJ
authors.lastname: Bray
authors.affiliation: Mater Medical Research Institute, South Brisbane, Queensland, Australia; School of Medicine, University of Queensland, St. Lucia, Queensland, Australia.
authors.forename: Celena F
authors.initials: CF
authors.lastname: Heazlewood
authors.affiliation: Mater Medical Research Institute, South Brisbane, Queensland, Australia.
authors.forename: David J
authors.initials: DJ
authors.lastname: Munster
authors.affiliation: Science and Engineering Faculty, Queensland University of Technology, Brisbane, Queensland, Australia; Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove, Queensland, Australia.
authors.forename: Dietmar W
authors.initials: DW
authors.lastname: Hutmacher
authors.affiliation: School of Medicine, University of Queensland, St. Lucia, Queensland, Australia; Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove, Queensland, Australia.
authors.forename: Kerry
authors.initials: K
authors.lastname: Atkinson
authors.affiliation: Queensland Eye Institute, South Brisbane, Queensland, Australia; School of Biomedical Sciences, Faculty of Health, Queensland University of Technology, Brisbane, Queensland, Australia; Institute of Health and Biomedical Innovation, Queensland University of Technology, Kelvin Grove, Queensland, Australia.
authors.forename: Damien G
authors.initials: DG
authors.lastname: Harkin
mesh_terms: D000818:Animals; D049109:Cell Proliferation; D064987:Cell- and Tissue-Based Therapy; D003315:Cornea / Q000276:immunology / Q000473:pathology*; D016039:Corneal Transplantation; D006801:Humans; D007165:Immunosuppression Therapy; D016850:Limbus Corneae / Q000166:cytology* / Q000276:immunology; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D011817:Rabbits; D013601:T-Lymphocytes / Q000166:cytology / Q000276:immunology
keywords: cell therapy; corneal limbus; immunosuppression; mesenchymal stromal cells